GINS MOTION REVEALS REPLICATION FORK PROGRESSION IS REMARKABLY UNIFORM

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 先前的研究已经导致了这样一幅画面，其中DNA的复制在芽殖酵母基因组的不同部分以可变的速率进行。这些先前的实验，专注于新生DNA的产生，已被解释为暗示复制叉进展的动力学受到局部染色质结构/架构的强烈影响，以及与控制转录、修复和表观遗传维持的机制的相互作用。在这里，我们采用了一种互补的方法来测定复制动态使用全基因组时间分辨染色质免疫沉淀结合微阵列分析的GINS复合物，复制叉的一个组成部分。令人惊讶的是，我们的数据表明，无论基因组位置如何，这种复合物都以高度均匀的速率进展，这表明酵母中的复制叉动力学比以前设想的更简单，更均匀。此外，我们展示了如何协同使用实验和建模导致新的生物学见解。特别是，一个简约的模型使我们能够准确地模拟整个基因组的叉运动，也揭示了一个微妙的现象，我们解释为低频叉逮捕所产生的。 描述这些结果的手稿已经发表： M.D. Sekedat，D. Feny？，R.S.放大图片作者：A. Aitchison，B.T. Chait“GINS Motion Reveals Replication Fork Progression is Remarkably Uniform Throughout the Yeast Genome”Mol Systems Biol，6（2010）353

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Previous studies have led to a picture wherein the replication of DNA progresses at variable rates over different parts of the budding yeast genome. These prior experiments, focused on production of nascent DNA, have been interpreted to imply that the dynamics of replication fork progression are strongly affected by local chromatin structure/architecture, and by interaction with machineries controlling transcription, repair and epigenetic maintenance. Here, we adopted a complementary approach for assaying replication dynamics using whole genome time-resolved chromatin immunoprecipitation combined with microarray analysis of the GINS complex, an integral member of the replication fork. Surprisingly, our data show that this complex progresses at highly uniform rates regardless of genomic location, revealing that replication fork dynamics in yeast is simpler and more uniform than previously envisaged. In addition, we show how the synergistic use of experiment and modeling leads to novel biological insights. In particular, a parsimonious model allowed us to accurately simulate fork movement throughout the genome and also revealed a subtle phenomenon, which we interpret as arising from low-frequency fork arrest. A manuscript describing these results has been published: M.D. Sekedat, D. Feny¿, R.S. Rogers, A.J. Tackett, J.D. Aitchison, B.T. Chait "GINS Motion Reveals Replication Fork Progression is Remarkably Uniform Throughout the Yeast Genome" Mol Systems Biol, 6 (2010) 353